Nu, nu&#39;-methylene-bis-amides for the suppression of tumors



3,085,940 N,N-METHYLENE-BliS-AMTDES FUR THE SUPPRESSION F TUMORS Andrew S. Tonicnt'cilr, Tappan, N311, Stuart D. Willson,

Park Ridge, NJ., and Adolph W. Vogel, Peal River, N.Y., assignors to American Cyanarnid Company, New Yorl-i, N.Y., a corporation of Maine No Drawing. Filed May 9, 1960, Ser. No. 27,488 8 Claims. (Cl. 167--65) This invention relates to novel compositions of matter for the treatment of malignant growths in mammals.

In recent years there has been noted a greater incidence of tumor growth than heretofore. This has been ascribed. to various reasons including longer life span, the pressures of modern life, food additives, tobacco, etc. A need exists, therefore, for compositions which are destructive to tumorous cells while at the same time having comparatively little or no elfeet on normal cells. In the past many compositions have been tried including, for example, the nitrogen mustards and other chemicals. These compositions have not been very satisfactory because of undesirable and sometimes severe side eitects.

We have now discovered that compounds represented by the following general tormula, when the active ingredient of the novel compositions of matter of the present invention, are useful in causing the remission oftumors R;( JNH-CHiNI-I( B in which R and R are the same and are selected from the group consisting of the allyl and 2-haloethyl radicals. The most efiective compounds in causing the remission of tumors are those according to the above general for mula wherein R and R are the same and are selected from the group consisting of the allyl, 2-bromoethyl, and 2-iodoethyl radicals.

The compounds of the present invention may be readily prepared by the interaction of 2 moleslof B-buteneamide or 2 moles of the appropriate '3-halopropionamide with 1 mole of formalhyde in the presence of a strong acid. Ordinarily, the reaction is carried out for a period of time ranging from a 'few minutes to an hour or more and at a temperature ranging from 30 C. to 100 C. Alternatively, the appropriate nitrile may be substituted for the 3-buteneamide or 3-halopropionamide as the starting material. In such case, hydrolytic conditions are employed so that the nitrile is hydrolyzed to its corresponding amide. Furthermore, alternate forms of formaldehyde such as paraformaldehyde may also be used. Generally, the desired product may then be recovered from the reaction mixture by concentration or by dilution with water.

The compounds of the present invention were tested against the tumors Sarcoma 180, lymphosarcoma 6C HED and C H mammary adencarinoma 72 Tumors from donor mice were minced and the particles transplanted, using a trocar, and inserted subcutaneously into the axillary region of a C l-I inbred strain of mice. When the tumors had grown to a palpable size (5-14 days) depending on the particular tumor, the test mice were grouped according to tumor size and treatment was started. An exception was the lymphosarcoma 6C HED in which treatment was started 2 to 3 days after implant before tumors were palpable. Treatment was by daily intraperitoneal injection for 6 days. Tumor harvest was on the seventh day following the first treatment. The tumors were weighed after excision and the following activity ratio was determined.

3,085,940 Patented Apr. 16, 1963 Activity ratio Average Weight oftumors from control mice -Average weight of tumors from treated mice A compound was judged active if its activity ratio exceeded 3.50in the case of Sarcoma 180, 6.39 in the case of lymphosarcoma 6C HED, and 3.72 in the case of adenocarcinoma 72j..

The following table summarizes the testing results for The dosage of the compounds of the present invention differs according to the compound, and the progression of the disease. The oral dose per day may vary from 5 to 500 mg. per kg. of body weight. A divided dose may vary from 50 mg. to 5.0 g. and the frequency of administration may vary widely.

The activity of the compounds of the present inventionagainst animal tumors presages a usefulness in the clinical remission of lymphomas and leukemia-s for periods of time that will vary with -difierent individuals. However, the effectiveness o-tthe compounds of the present inven: tion in causing the remission of tumors in human subjects has not yet been demonstrated.

The compounds of the present invention may be orally administered in any. of the usual dosage unit forms of pharmaceutical preparations. These may take the form of tablets, capsules, pills, powders orany other desirable form in the therapeutic quantities set forth above. The dosage form may be for a single daily therapeutic dose or in smaller units for multiple doses or in larger units for division into single doses. It is understood that in addition to the therapeutic compound there may be present excipients, binders, fillers, and other therapeutically inert ingredients necessary in the formulation of pharmaceutical preparations.

The compounds of the present invention may also be administered parenterally by dissolving or suspending the compound in a parenterally suitable vehicle such as, for example, propylene glycol or polyethylene glycol, or by dissolving or suspending the compound in an aqueous solution of such vehicle.

The following examples show the preparation and formulation of the therapeutic compounds of the present invention.

Example 1 N, N'-METHYLENE-BIS-(s-BROMOPROPIONAMIDE) A solution of 8.3 g. of 37% tormaldehyde in 21 g. of concentrated sulfuric acid was added to 26.8 g. of 3- bromopropionit-rile at 35-40 C. After five hours the solution was poured onto ice and the crude product was removed by filtration. There was obtained 25 .0 g. (79% crude yield) of N,N-methylene-bis-(3-bromopropionamide). M.P. 184-186.5 C. Recrystallization from 80% acetone gave white crystals, M.P. 184.5-1-86.5 C.

3 Example 2 N,N -i\HETHYLENE-BIS- 3-IODOPROPIONA1MIDE) A mixture of 10.0 g. of 3-iodopropionamide, 0.75 g. of paraformaldehyde, and 0.8 ml. of concentrated hydrochloric acid in 150 ml. of ethylene dichloride was refluxed for minutes. There was obtained 9.4 g. (91% crude yield) of N,N-methylene-bis-(3-iodopropionamide), M.P. 191-193 C. dec. Recrystallization from 90% isopropyl alcohol gave 8.6 g. of white crystals, M.P. 193l94 C. dec.

Example 3 N,N' -METHYLENE-BIS- 3-BUTENEAMIDE) A solution of 8.1 g. of 37% formaldehyde in 21 g. of concentrated sulfuric acid was added dropwise to 13.4 g. of 3-butenenitrile at 15 C. The resulting thick syrup was stored overnight at 25 C. and then poured onto ice. The crude product was removed by filtration whereby there was obtained 10.7 g. (58% crude yield) of N,N'- methylene-bis-(3-buteneamide), M.P. 193-198 C. After recrystallization from 95% ethanol, there was obtained 8.6 g. of white crystals, M.P. 198199 C.

Example 4 A mixture of parts of N,N-methylene bis-(3- bromopropionamide), 15 parts of starch, and 1 part of magnesium stearate was thoroughly blended and then screened through a 60 mesh screen. The resulting powder was tableted in an automatic tableting machine whereby suitable 500 mg. scored white tablets were obtained.

What is claimed is:

1. A therapeutic composition for oral administration useful for the remission of tumors in dosage unit form comprising from 0.25 g. to 50 g. of a compound having the general formula 0 R.( i-NHoHr-NH- iRl wherein R and R are the same and are selected from the group consisting of the allyl and 2-haloethyl radicals, and a solid pharmaceutical carrier.

2. A therapeutic composition for oral administration useful for the remission of tumors in dosage unit form comprising from 0.25 g. to 50 g. of N,N-methylene-bis- (3-bromopropionamide) and a solid pharmaceutical carrier.

3. A therapeutic composition for oral administration useful for the remission of tumors in dosage unit form comprising from 0.25 g. to g. of N,N'-methylene-bis- (3-iodopropionamide) and a solid pharmaceutical carrier.

4. A therapeutic composition for oral administration useful for the remission of tumors in dosage unit form comprising from 0.25 g. to 50 g. of N,N'-methylene-bis- (3-butenearnide) and a solid pharmaceutical carrier.

5. A method for causing the remission of tumors in mammals which comprises administering in dosage unit form from 5 mg. to 500 mg. per kg. of body weight per day of a compound of the formula:

R1 iNH-0HzNH-( i-R2 wherein R and R are the same and are selected from the group consisting of the allyl and 2-haloethyl radicals.

6. A method for causing the remission of tumors in mammals which comprises administering in dosage unit form 'from 5 mg. to 500 mg. per kg. of body weight per day of N,N-methylene-bis-(3-bromopropionamide).

7. A method for causing the remisison of tumors in mammals which comprises administering in dosage unit form from 5 mg. to 500 mg. per kg. of body weight per day of N,N-methylene-bis-(3-iodopropionamide) 8. A method for causing the remission of tumors in mammals which comprises administering in dosage unit form from 5 mg. to 500 mg. per kg. of body weight per day of N,N-methylene-bis-(3-buteneamide).

References Cited in the file of this patent UNITED STATES PATENTS 2,475,846 Lundberg July 12, 1949 FOREIGN PATENTS 743,466 Germany Nov. 4, 1952 OTHER REFERENCES Pfizer Spectrum, Screening of Anti-Cancer Agents, vol. 7, No. 3, March 1959, pp. 62-63.

American Jurisprudence-Proof of Facts, Annotated, vol. 3, Brakes to Damages, 1959 by Jurisprudence Publishers, Inc., The Lawyers Co-operative Publishing Co., Rochester, N.Y., Cancer, pp. -131. 

1. A THERAPEUTIC COMPOSITION FOR ORAL ADMINISTRATION USEFUL FOR THE REMISSION OF TUMORS IN DOSAGE UNIT FORM COMPRISING FROM 0,25 G. TO 50 G. OF A COMPOUND HAVING THE GENERAL FORMULA 